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Scepter™ 2.0 Cell CounterPrecise, handheld cell counting

Product Brochure

EMD Millipore is a division of Merck KGaA, Darmstadt, Germany

http://www.millipore.com/?cid=BIOS-S-EPDF-1028-1202-DS

Trust Scepter™ counting with your most valuable samples to get reproducible and reliable counts. The reliability of Scepter™ counting is particularly apparent with smaller cell types. Because the Scepter™ cell counter measures volume using the Coulter Principle, it can quantify cells based on size and will discriminate larger cells from smaller debris, unlike vision based techniques, which rely on object recognition software and cannot reliably detect small cells.

Scepter™ 2.0 is your portable cell counter. While other automated counters consume bench space and rely on object recognition software, manual focusing, and clumsy loading chambers, the Scepter™ cell counter provides true automation without the error that accompanies vision-based systems. With its microfabricated, precision- engineered sensor, the Scepter™ cell counter does all the work and delivers accurate and reliable cell counts in less than 30 seconds.

“At last, an alternative to lining up for the Coulter Counter ®, and far easier than sweating over fragile hemocytometers.”

AMY A. CAUDY is a Lewis-Sigler Fellow at Princeton University’s Lewis-Sigler Institute for Integrative GenomicsThe Scientist, Dec. 2010. Top Ten Innovations of 2010.

“Cell counting is normally a very tedious process and usually only provides minimal information on the cell population. This instrument, which is only slightly larger than an automatic pipette, allows you to count cells in your tissue-culture hood, simplifies the procedure, and provides much useful data, such as the fraction of intact cells.”

H. STEVEN WILEY is a lead biologist at the Environmental Molecular Sciences Laboratory at the Pacific Northwest National LaboratoryThe Scientist, Dec. 2010. Top Ten Innovations of 2010.

What people are saying...

Scepter™ 2.0 – Precise, handheld cell counting

The power of precision

Scepter™ 2.0 marks the next generation in Scepter™ technology, highlighted by:

Compatibility with More Cell Types The Scepter™ cell counter is the only one on the market to accurately count particles as small as 3 µm in diameter

Increased Cell Concentration Range The new 40 µm sensor can count samples with concentrations as high as 1,500,000 cells/mL

Powerful Software for Complex, Effortless Cell Analysis

• Compare sample sets side by side using histogram overlay and multiparametric data table

• Create and save gating templates

• Generate reports, graphs and tables

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Scepter™ sensor technologyCompatible with 60 µm and 40 µm sensors, the Scepter™ 2.0 cell counter can meet even more of your cell- and particle-counting needs. Use the 60 µm sensor for particles between 6 and 36 µm. Use the 40 µm sensor for particles between 3 and 17 µm.

• Precise volumes are drawn into the Scepter™ sensor.

• As cells flow through the aperture in the sensor, resistance increases. This increase in resistance causes a subsequent increase in voltage.

• Voltage changes are recorded as spikes with each passing cell.

• Spikes of the same size are bucketed into a histogram and counted. This histogram gives you quantitative data on cell morphology that can be used to examine the quality and health of your cell culture.

+ -

Figure 1. Particles are detected by Ohm’s Law V=IR(V=voltage, I=current, and R=resistance)

Table 1. Cell types validated with the Scepter™ cell counter and the recommended Scepter™ sensor.

Cell TypeMeasured size (µm)

40 µm sensor

60 µm sensor

2102 Ep 15-19454 beads A172 15A253 14-18A375 16A431 15-17A549Algae (various) 7-9B35 13-16B Cells 6-11C2C12 12C305 12-14C6 12-13CA46 10-12Caco-2 17CHO 14-17COS-1 12Cos-7 15D283 12Daudi 10-12DU-145 15-17Epithelia 14-15HCT-116 10HEK293 11-15HeLa 12-14HepG2 12HFF 18-20Hs27 14HT-1080 14-16HT-29 11HUH7- Hepatoma line

Human ES Cells 9-12HUVEC 14-15IMR-32 12-14IMR-90 15Jurkat 13K562 22KB 14KG-1 10-13L6 14-16LNCaP 15-16Luminex® beads 5-6MCF7 15-17MDCK 13-15

Cell TypeMeasured size (µm)

40 µm sensor

60 µm sensor

Meg-01 16-17MG-63 15-17Mouse ES Cell 5-13Mesenchymal Stem Cell 15-16 MRC-5NCI-H146 10-13NIH 3T3 15NTERA2, clone D1 13OK 17-18PBMCs 7-12PC12 9-13Primary Astrocytes 7Primary Neuronal Cell Raji 12-15Ramos 11-12Rat Dorsal Root Ganglion Cells

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Rat Whole Blood 4.6Red Blood Cells 5-7Rat Neural Stem Cell 11-13RAW 264.7 12-15RBL 11-13RIN-mF5 13-14SF9 13SH-SY5Y 12Sk-Br-3 15-20SK-MEL-28 17-19SK-N-MC 14-15SK-N-SH 14-15Splenocytes 7-9SW-480 15SW-620 13-14T84 14-18T98G 17TF-1 13-14U251 16-20U2OS 16-19U266 12U87-Human Glioblastoma cell line

12-14

U937 11-13WI-38 12-15Y79 13-14Yeast- Pichia Pastoris 5Yeast- S.cerevisiae 6

n Recommended based on size n EMD Millipore Validated n Customer Validated

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Scepter™ counting delivers precision

Figure 2. The average percent coefficient of variation (CV) for each counting method shown was calculated using cell concentration measurements at 50,000 cells/mL samples of 19 different cell lines. The Scepter™ cell counter is more precise than vision-based counting and hemocytometry, and approaches the precision of the Coulter Counter® standard (maroon bars). Error bars represent standard deviation.

No camptothecin 100 µM camptothecin

1009080706050403020100

MDMA231 cells

Flow cytometry


Scepter™ Trypan blue + hemocytometerFlow cytometry Scepter™ Trypan blue + hemocytometer

1009080706050403020100

NIH 3T3 cells

60.00%

50.00%

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80.00%

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Perc

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% V

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% V

iabi

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Scepter™ cell counter Vision-based counter Hemocytometer

= Average % CV = Coulter counter® % CV

Format Counting methods

Sample volume needed

Sample volume counted

Cells counted in a 100,000 cell/mL sample

Average % CV

Hemocytometer Slide and microscope

Manual, vision-based 10 µL .1 µL /square

10/square 41.8

Brand L Benchtop Automated vision-based system

10 µL .4 µL 40 32.1

Scepter™ Cell Counter Handheld Impedance-based cell detection 100 µL 50 µL 5000 9.1

As easy as pipettingPrepare the sample:Start with a single-cell suspension, diluted to a total volume of 100 µL (recommended) in phosphate buffered saline (such as EmbryoMax® 1x DPBS) to 10,000-500,000 cells/mL (operating range for 60 µm sensor) in a 1.5 mL microcentrifuge tube.

Perform cell count:• Turn on the Scepter™ cytometer by pressing the

toggle on the back of the instrument and wait for on-screen instructions to appear.

• When prompted, attach a sensor to the end of the Scepter™ unit with the electrode sensing panel facing toward the front of the instrument, and you’ll see detailed instructions for each step of the counting process.

• Pipette once to draw sample into the sensor. 50 µL of your cell suspension is drawn into the microfabricated, precision-engineered channel embedded in the sensor. The cell sensing zone detects each cell drawn into the sensor and thus cell concentration is calculated.

• The sensing zone also measures cell sizes and cell volumes with sub-micron and sub-picoliter resolution, enabling the Scepter™ cytometer to display a histogram distribution of cell size or cell volume.

There is no need to subjectively determine cell counts, as required by vision-based counting methods. The Scepter™ cell counter detects every cell and displays the population as a histogram of cell size distributions. From the histogram, count all the cells or use the gating function to count a chosen subpopulation. By monitoring changes in your histogram, you can gain insight into the health and quality of your cell culture from one experiment to the next.

3 4

2

Average cell volume (pL)

Average cell diameter (µm)

Cell concentra-tion (cells/mL)Histogram displayed as

function of cell diameter or cell volume

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A VIEW OF SCEPTER™ SOFTWARE PRO DATA: data files from your Scepter™ cell counter

CURRENT PLOT: working plot and data file

MULTIFUNCTIONAL PLOT: multiple data sets/histogram overlays

ANALYSIS TEMPLATES: saved gating parameters

GROUP STATS: customizable statistics from your selected data files

REPORTS: export, print selected graphs/files, cut and paste

Intuitive analysis softwareFrom simple counts to complex volume measurements used to assess cell health parameters, Scepter™ Software Pro provides an intuitive, intelligent platform to perform high-level cell analysis based on the size measurements captured with the Scepter™ cell counter.

Using the Scepter™ Software Pro on your computer, you can:• Compare several samples

and data sets side by side using histogram overlay and multipara-metric tables

• Create and save gates to be used from one experiment to the next

• Create attractive graphical presentations and reports with your data

1

4 5 6 7

2 3

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Scepter™ 2.0 for cell health Instantly gauge the health of your cell cultures without even leaving the culture hood. Because the Scepter™ cell counter displays high-resolution histograms of entire cell populations, you can differentiate live cells from dead cells and debris by simply gating on the histogram peak corresponding to larger-diameter cells. No staining is required! The resulting calculation for % viable cells agrees with viability calculations obtained using flow cytometry (ViaCount® reagent) and Trypan blue staining/hemocy-tometry (shown here with MDMA231 and NIH 3T3 cells).

Scepter™ 2.0 for counting heterogeneous cell populations Count blood cells and other cells with small diameters with the highest precision. Biological samples such as primary isolates or cultured cells are often heterogeneous mixtures of cells that differ by type and/or function. Such differences in cellular attributes are most commonly determined by multicolor fluorescent antibody detection of cell type specific surface marker(s) using flow cytometry. Notably, in addition to variations in protein expression, many cell types and physiological states are also uniquely distinguishable on the basis of size alone. The ability to identify population subsets on the basis of phenotypic differences and further determine their relative frequencies (and concentrations) is critical to many aspects of research.

Distinguishing lymphocytes from monocytes in freshly isolated PBMCs. The assessment of immune profiles of the various immune cell subsets can help identify molecular signatures that may facilitate research. The Scepter™ cell counter, when used in combination with Scepter™ Software Pro, provides a tool for rapid determination of lymphocyte and monocyte concentrations as well as the relative frequency of these cell types in PBMC isolates.

Figure 3. Rapid cell analysis using the Scepter™ 2.0 device provides reliable assessments of cell viability compared to flow cytometry (ViaCount® assay) and hemocytometry (using Trypan blue staining). MDMA231 cells (A) and NIH 3T3 cells (B) were treated with camptothecin 24 hours prior to analysis.

Figure 4. The Scepter™ 2.0 cell counter counts PBMCs with greater precision than other counting methods, as reflected by low coefficients of variation. % CVs were calculated using average cell counts of four replicate samples.

3025

% C

oeffi

cien

t of

Var

iatio

n (%

CV)

Scepter Counter(40 µm sensor)

Automated Vision-based Counter

Hemocytometer

COUNTING METHOD

201510

50

20

15

% C

oeffi

cien

t of

Var

iatio

n (%

CV)

Scepter Counter(40 µm sensor)

Average %CVZ2 Coulter Counter

RBCs

PBMCs

Automated Vision-based Counter

Hemocytometer

Counting Method

20

5

0

Average % CV Z2 Coulter Counter®

APPLICATIONS


1009080706050403020100

MDMA231 cells

Flow cytometry



1009080706050403020100

NIH 3T3 cells

60.00%

50.00%

70.00%

80.00%

40.00%

30.00%

20.00%

10.00%

0.00%

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V

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1009080706050403020100

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1009080706050403020100

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20.00%

10.00%

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A.

B.

EMD Millipore continues to expand the capabilities of Scepter™ technology. And the latest generation, Scepter™ 2.0, features enhanced analytical powers, enabling you to count even more cell types and sizes.

7

Test Cell Fraction Scepter™1Forward Scatter2 Staining3

1 Lymphocyte 58 65 63

Monocyte 42 35 37


Monocyte 32 28 29


Monocyte 34 31 29

Coun

t

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Plot PO3, ungated Current Acquisition

Diameter (µm)

00 1000 2000 3000 4000 5000 6000 7000 8000 9000 10000

102030405060708090

100

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Flow Cytometry Scepter™ (40 µm)Figure 5. Representative comparison of histogram plots for human PBMC samples acquired on the Scepter™ cell counter (diameter histogram on right) and guava easyCyte™ flow cytometry (forward scatter histogram on left) platforms. Analysis plots derived from both platforms demonstrate three distinct peaks corresponding to 1) dead cell/debris, 2) lymphocyte and 3) monocyte fractions. The difference in counts displayed (Y-axis) is due to differences in sample dilution between the guava easyCyte™ flow cytometer and the Scepter™ cell counter.

Table 2. Lymphocyte and monocyte subset frequencies from three individual PBMC samples. Aliquots from each sample were analyzed using the guava easyCyte™ flow cytometry and Scepter™ platforms. 1Values were derived from the diameter histogram plot. 2Values were derived from the forward scatter histogram plot based on total events measured on guava easyCyte™ flow cytometry platform. 3Staining frequencies derived as follows: % Lymphocytes = (% CD3+ T cells) + (%CD16/56+ NK cells) + (%CD19+ B cells); % Monocytes = % CD14+ cells

Precise and accurate bead counting with Scepter™ 2.0Micron-sized beads are used in a variety of biologi-cal applications, ranging from daily validation of flow cytometer performance to purification of fusion protein constructs from cell lysates. Accurate determination of bead counts at the onset of each assay allows for standardization of bead concentrations across multiple samples and minimizes errors and variation in down-stream results. The Scepter™ cell counter is well suited for precise counting for beads of numerous types and can improve reproducibility of bead-based assays, such as immunoprecipitation and multiplexed detection.

Scepter™ counting can facilitate yeast cell counting for brewing and wine industriesIntroduction of a consistent yeast cell concentration is required for successful beer and wine fermentation as well as to maintain batch-to-batch reproducibility and ensures consistent fermentation over many cycles. Scepter™ counting can be used to monitor yeast size and concentration by yielding interpretable histograms that could be gated to provide this depth of information.

Figure 6. Scepter™ Software Pro displays imported size distribution histograms as either a single sample histogram or as overlaid histograms for multiple samples. Shown is an overlaid histogram for serially diluted 5.6 µm MILLIPLEX® map microspheres.

Figure 7. The Scepter™ cell counter counts yeast cells with good accuracy and linearity. Measured yeast cell concentrations were compared to theoretical concentrations. The solid gray line represents the theoretical values. Dotted lines represent best linear fit to data. Both the Scepter™ and Coulter Counter® platforms show a loss of linearity and accuracy upon an increase in cell concentration.

0

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2000

3000

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5.6 µm Polystyrene MILLIPLEX® MAP Antibody-Conjugated Microspheres

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t

Diameter (µm)3 3 9 12 15 18

1,500,000750,000500,000250,000125,00050,000

0.0E+000.0E+00

4.0E+05

8.0E+05

Mea

sure

d Ce

ll Co

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Theoretical Cell Concentration(cells/mL)

1.2E+06

1.6E+06

4.0E+05 8.0E+05 1.2E+06 1.6E+06

R2=0.9983

R2=0.9953

Theoretical Cell ConcentrationZ2 Coulter Counter®Scepter™ - 40 µm Sensors

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Ordering InformationDescription Qty Catalogue No.

Scepter™ 2.0 Handheld Automated Cell Counter

with 40 µm Scepter™ Sensors (50 Pack) 1 PHCC20040

with 60 µm Scepter™ Sensors (50 Pack) 1 PHCC20060

Includes:

Scepter™ Cell Counter 1

Downloadable Scepter™ Software 1

O-Rings 2

Scepter™ Test Beads 1 PHCCBEADS

Scepter™ USB Cable 1 PHCCCABLE

Scepter™ Sensors, 60 µm 50 PHCC60050

500 PHCC60500

Scepter™ Sensors, 40 µm 50 PHCC40050

500 PHCC40500

Universal Power Adapter 1 PHCCP0WER

Scepter™ O-Ring Kit, includes 2 O-rings and 1 filter cover 1 PHCC0CLIP

EmbryoMax, MILLIPLEX and ViaCount are registered trademarks of Merck KGaA, Darmstadt, Germany. EMD Millipore, the M mark, guava easyCyte and Scepter are trademarks of Merck KGaA, Darmstadt, Germany. Coulter Counter is a registered trademark of Beckman Coulter. Luminex is a registered trademark of Luminex Corporation.Lit No. PB2217EN00 Fisher Lit No. BN1124111 LS-SBU-11-05340 02/2012 Printed in the USA. © 2012 EMD Millipore Corporation, Billerica, MA, USA. All rights reserved.

Are you an existing Scepter™ user interested in

upgrading your device to Scepter™ 2.0? It’s easy.

Visit www.millipore.com/scepterupgradeto upgrade your Scepter™ today!

For technical assistance, contact Millipore:1-800-MILLIPORE (1-800-645-5476)E-mail: [email protected]

http://www.millipore.com/catalogue/item/PHCC20040&cid=BIOS-S-EPDF-1028-1202-DS


http://www.millipore.com/catalogue/item/PHCCBEADS&cid=BIOS-S-EPDF-1028-1202-DS

http://www.millipore.com/catalogue/item/PHCCCABLE&cid=BIOS-S-EPDF-1028-1202-DS





http://www.millipore.com/catalogue/item/PHCCP0WER&cid=BIOS-S-EPDF-1028-1202-DS

http://www.millipore.com/catalogue/item/PHCC0CLIP&cid=BIOS-S-EPDF-1028-1202-DS

http://www.millipore.com/life_sciences/flx4/scepter_muse_register&cid=BIOS-S-EPDF-1028-1202-DS

http://www.millipore.com/?cid=BIOS-S-EPDF-1028-1202-DS

Scepter 2 Product Brochure - Fisher Scientificfscimage.fishersci.com/.../scepter_brochure.pdf ·...

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Transcript of Scepter 2 Product Brochure - Fisher Scientificfscimage.fishersci.com/.../scepter_brochure.pdf ·...